The present invention relates to a reaction-measurement apparatus for observation and examination of specimens such as cytodiagnosis or histopathological samples using a microscope etc. More particularly, this invention concerns an apparatus and method in which a reagent is made to react quickly in an area of special notice and its results can be quickly measured.
Examinations such as pathological examination and cytodiagnosis are important in examination for cancer, etc. In this kind of examination, pieces of tissue, cell, etc. are usually removed and immobilized by smearing them on the slide glass to prepare specimens. After such treatments as staining, the specimens are examined and diagnosed. Another method in which infection or abnormality is detected by allowing a cell to react with a fluorescent antibody marker or nucleic acid probe is widely used for cytodiagnosis and examination for infectious disease (Japanese Patent Laid-open No. 2000-310637, and “A Clinical Examination Special Issue <Cytodiagnosis—Prospects in the 21st century > Vol. 44, No. 11” published by IGAKU-SHOIN, LTD. Tokyo, October 2000).
A variety of staining techniques are known. Papanicolaou stain, Hematoxylin-eosin stain, Giemsa stain, etc. are used depending on the purpose. In Papanicolaou stain, for example, the cell is stained in the following procedure and used for cytodiagnosis: nuclear staining with Gill's Hematoxylin; rinsing with water and ethanol; decoloring and fractionation with alcohol hydrochloride; rinsing; coloring of nucleus with ammonia alcohol; rinsing; staining of cytoplasm; rinsing; dehydration; and penetration. An automation apparatus for staining the cell immobilized on such a slide glass has already been put on the market. An example of such an apparatus is disclosed in unexamined Japanese Patent Laid-open No. 62-27682, etc.
In recent years, in addition to the above-mentioned cytodiagnosis, there is developed a method using an oligonucleotide probe or DNA probe which hybridizes with concerned virus DNA sequence etc. of virus gene, cancer-related gene etc. to directly find whether the gene concerned is present in the tissue or cell. By using the oligonucleotide probe, DNA probe or the like in which a fluorescent substance, light-emitting substance etc., DNA probe etc. are marked, it is possible to grasp a gene present in the specimen by hybridizing and to detects it in fluorescent or light-emitting measurement, etc.
In cytodiagnosis etc., it is judged whether there are abnormalities by checking the form of cell, the size of the nucleus-cytoplasm, relations between cells.
The oligonucleotide probe, DNA probe, etc. can quantitatively detect the presence of objective genes DNA or RNA. The presence of a gene is important information in that the presence of a gene is a risk factor indicating that disease can develop. In virus infection, it can be found out before the symptoms of the disease are shown, and thus more effective treatment for the disease will become possible. Accordingly, a reliable diagnosis will be possible by combining the detection of a gene by the oligonucleotide probe or DNA probe, etc. and the measurement of the conventional cytodiagnosis.
Generally, a gene in the cell is detected through fluorescent detection by hybridizing fluorescent marker oligonucleotide probe, DNA probe, etc. Furthermore, the conventional cytodiagnosis is performed by measuring a transmitted light image after the usual staining described above. The hybridizing and the usual staining are different in procedure and are not conducted at the same time but individually.
Generally, hybridizing reaction needs a long time, several hours to one night, and therefore is poor in operability. It takes one or more hours to immune-stain a specimen on slide glass. Furthermore, a large quantity of reagent is required, since the reagent is applied all over the specimen portion on the slide glass, not on a limited or necessary portion. In particular, the reagent for hybridization reaction is costly.